Primer, kit and method for detecting Dendroctonus pseudotsugae Hopkins
A technology of beetles and kits, applied in the field of molecular biology, can solve the problems of difficult identification, rapid identification, high personnel requirements, etc., and achieve good specificity
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[0046] The nested PCR detection method of embodiment 1 Huangshan beetle
[0047] (1) Reagents used: EZgene Insect gDNA Kit kit: purchased from BIOMIGA Company; SYBR PremixEx TaqⅡ (Perfect Real time): purchased from Bao Bioengineering (Dalian) Co., Ltd.; DL2000 DNA Marker: purchased from Bao Bioengineering (Dalian) Co., Ltd.; 1×Loading Buffer: purchased from Bao Biological Engineering (Dalian) Co., Ltd.
[0048] (2) DNA extraction: use the EZgene Insect gDNA Kit kit from BIOMIGA Company to extract target insects (including Beetles pine-toothed beetles, Beetles largii, Beetles largii, Beetles punctatus and Beetles pine). ) gDNA, dilute the DNA to 50ng / μL.
[0049] (3) Using gDNA as a template, the first round of PCR amplification of the COI sequence was performed using universal primers (Table 1-1), with a fragment size of 700 bp.
[0050] Table 1 Sequences of general primers LCO1490 and HCO2198
[0051]
[0052] The PCR reaction system is shown in Table 2.
[0053] Table...
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